Molecular identification, phylogeny and antifungal susceptibilities of dematiaceous fungi isolated from human keratomycosis.

PURPOSE: To investigate the dematiaceous fungal profile of patients with ocular mycoses attending a tertiary eye care hospital in Coimbatore, India METHODS: The identification of dematiaceous fungus based on their morphology, their genotypes, and the measurement of the minimum inhibitory concentrations (MICs) using microdilution method of routinely used antifungal drugs were all compared. RESULTS: A total of 148 dematiaceous fungi were isolated during a study period of 27 months. Isolates were confirmed as Curvularia spp. (n = 98), Exserohilum spp. (n = 32), Alternaria spp. (n = 14), Exophiala spp. (n = 2), Cladosporium sp. (n = 1) and Aureobasidium sp. (n = 1). Out of 50 well grown isolates characterized genotypically based on the amplification and sequencing of the ITS region of the ribosomal RNA gene cluster and subsequent BLAST analysis, Curvularia lunata (n = 24), C. aeria (n = 1), C. spicifera (n = 8), C. hawaiiensis (n = 1), C. maydis (n = 2), C. papendorfii (n = 2), C. geniculata (n = 3), C. tetramera (n = 2) and Exs. rostratum (n = 7) were identified. In vitro antifungal susceptibilities of the most tested dematiaceous isolates showed that voriconazole had a MIC50 of 0.25 µg ml-1, while amphotericin B had a MIC50 of 0.25 µg ml-1 for Curvularia spp. and Alternaria spp. CONCLUSION: Voriconazole proved to be the most effective drug against the pigmented filamentous fungi, followed by amphotericin B, itraconazole and econazole.

In-vitro assessment of first-line antifungal drugs against Aspergillus spp. caused human keratomycoses.

BACKGROUND AND OBJECTIVES: Aspergillus keratitis are in the increasing trend and reported as the second most common cause of mycotic keratitis in developing countries. The present study was designed to isolate, identify Aspergillus spp. from the keratits/corneal ulcer patients attending a tertiary care eye hospital, Coimbatore, South India and to assess the minimum inhibitory concentrations (MICs) against ten clinically used first-line antifungal drugs. METHODS: A total of seventy-three Aspergillus strains isolated from corneal scrapings were included and assessed for a period of one year. All isolates were identified up to the species level by morphological observations. Antifungal drug susceptibilities were determined against a standard panel of antifungal agents. CONCLUSIONS: Five different species of aspergilli, A. flavus (n=53), A. fumigatus (n=14), A. terreus (n=9), A. tamarii (n=6) and A. niger (n=3) were identified based on morphological features. Minimum inhibitory concentration analyses indicated that, voriconazole, natamycin, itraconazole, clotrimazole, econazole followed by ketoconazole shall be the order of choices for the effective treatment for Aspergillus keratitis.